Measurement of Cholesterol Transfer from Lysosome to Peroxisome Using an In Vitro Reconstitution Assay

• DOI: 10.1007/978-1-4939-6875-6_11
• Publication Date: 2017-02-15T03:57:36Z
• Authors: Jie Luo; Ya-Cheng Liao; Jian Xiao; Bao-Liang Song
• Keywords: Cholesterol; Peroxisomes; Biological Transport, Active; Humans; Intracellular Membranes; Lysosomes; HeLa Cells

Low-density lipoproteins (LDLs) are taken up by the cell mainly through receptor-mediated endocytosis. LDL-derived cholesterol leaves lysosome and further transports to downstream organelles for specific cellular needs. We recently report that cholesterol transfers from lysosome to peroxisome through lysosome-peroxisome membrane contact (LPMC). Here, we use iodixanol density gradient centrifugation to isolate lysosomes and peroxisomes separately for the in vitro reconstitution of LPMC. We also apply 3H-cholesterol-labeled lysosomes and peroxisomes in vitro to measure 3H-cholesterol transfer through LPMC.